Quantitative Analysis of Tissue Distribution of the B16BL6-Derived Exosomes Using a Streptavidin-Lactadherin Fusion Protein and Iodine-125-Labeled Biotin Derivative After Intravenous Injection in Mice
| Autor: | Masaki Morishita, Takafumi Imai, Kana Kato, Yoshinobu Takakura, Yuki Takahashi, Hideo Saji, Kohei Sano, Makiya Nishikawa, Takuma Yamashita |
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| Rok vydání: | 2015 |
| Předmět: |
Male
Streptavidin Recombinant Fusion Proteins Melanoma Experimental Biotin Pharmaceutical Science Biology Exosomes Exosome Iodine Radioisotopes Mice chemistry.chemical_compound Gaussia Animals Tissue Distribution Luciferase Lactadherin Mice Inbred BALB C biology.organism_classification Molecular biology Fusion protein Microvesicles chemistry Injections Intravenous Protein Binding |
| Zdroj: | Journal of Pharmaceutical Sciences. 104:705-713 |
| ISSN: | 0022-3549 |
| Popis: | We previously succeeded in the visualization of tissue distribution of B16BL6 cells-derived exosomes by labeling with Gaussia luciferase (gLuc)-LA, a fusion protein of gLuc (a reporter protein) and lactadherin (LA; an exosome-tropic protein). However, total amount of B16BL6-derived exosomes delivered to each organ could not be evaluated because of the reduction of luminescent signal from gLuc-LA. The aim of the present study was to quantitatively evaluate the tissue distribution of B16BL6-derived exosomes. To this end, we labeled B16BL6-derived exosomes with iodine-125 ((125) I) based on streptavidin (SAV)-biotin system. A plasmid vector encoding fusion protein, SAV-LA, was constructed, and B16BL6 cells were transfected with the plasmid to obtain SAV-LA-coupled exosomes. SAV-LA-coupled exosomes were incubated with (3-(125) I-iodobenzoyl) norbiotinamide ((125) I-IBB) to obtain (125) I-labeled B16BL6 exosomes. After intravenous injection of (125) I-labeled B16BL6 exosomes into mice, radioactivity quickly disappeared from the blood circulation. At 4 h, 28%, 1.6%, and 7% of the injected radioactivity/organ was detected in the liver, spleen, and lung, respectively. These results indicate that (125) I-labeling of exosomes using SAV-biotin system is a useful method to quantitatively evaluate the amount of exogenously administered exosomes delivered to each organ and that the liver is the major organ in the clearance of exogenously administered B16BL6-derived exosomes. |
| Databáze: | OpenAIRE |
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